Journal: Science translational medicine
Article Title: An antibody-drug conjugate directed to the ALK receptor demonstrates efficacy in preclinical models of neuroblastoma
doi: 10.1126/scitranslmed.aau9732
Figure Lengend Snippet: Neuroblastoma cells expressing wild-type ALK (NB-1 and IMR-32) and mutated ALK (SK-N-SH) or with no detectable ALK expression (SK-N-AS) were treated with increasing doses of CDX-0125-TEI, a control IgG1 antibody conjugated with TEI (IgG1-TEI), or the free payload (NMS-P528). Titration of CDX-0125-TEI induced cytotoxic activity in all ALK-expressing models, with IC50 values in the picomolar range and independent of ALK mutation status or number of cell surface ALK receptors. By contrast, no measurable IC50 value could be derived in the ALK-negative cell line SK-N-AS. Free NMS-P528 elicited complete cell killing at sub-picomolar concentrations in all cell lines tested. Quoted IC50 and ALK expression values reflect calculated means and SEM (n = 3) from at least three independent experiments.
Article Snippet: Upon immobilization onto polyvinylidene difluoride (PVDF) membranes, proteins were labeled using one of the following antibodies: phosphorylated ALK (Y1278), total ALK, cleaved caspase-3, and GAPDH (Cell Signaling Technology).
Techniques: Expressing, Titration, Activity Assay, Mutagenesis, Derivative Assay